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PMID: 3039492 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Characterization of a mammalian mutant with a camptothecin-resistant DNA topoisomerase I.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 84 ·No. 16 ·1987-08-00 ·Pages 5565-9

Andoh T, Ishii K, Suzuki Y, Ikegami Y, Kusunoki Y, Takemoto Y, Okada K

Abstract

DNA topoisomerase I was purified to near homogeneity from a clonal line of human lymphoblastic leukemia cells, RPMI 8402, that is resistant to camptothecin, a cytotoxic alkaloid from Camptotheca acuminata, and compared with that of the parent wild-type cells. As assayed by relaxation of the supercoiled plasmid DNA and by formation of enzyme-linked DNA breaks, the purified enzyme from the resistant cells was shown to be greater than 125-fold as resistant to camptothecin as the wild-type enzyme, comparable to a cellular resistance index of about 300. Therefore, the cellular resistance appears to be due to the resistance of the enzyme. The amount of the immunoreactive enzyme protein in whole extract appeared to be reduced to less than half that of the wild-type enzyme. These results establish that DNA topoisomerase I is the cellular target of camptothecin and that DNA topoisomerase I is essential for the survival of mammalian cells.

MeSH Terms
Camptothecin/pharmacology Cell Line DNA Topoisomerases, Type I/metabolism DNA, Superhelical/drug effects Drug Resistance Humans Immunosorbent Techniques Leukemia, Lymphoid/enzymology Mutation Nucleic Acid Conformation/drug effects Peptide Mapping Plasmids
Chemicals
DNA, Superhelical DNA Topoisomerases, Type I Camptothecin
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Andoh T
Ishii K
Suzuki Y
Ikegami Y
Kusunoki Y
Takemoto Y
Okada K
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39 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.84.16.5565
Published
1987-08-00
Pages
5565-9
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC298903
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.84.16.5565 PMC Full Text View journal details
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